Bioinformatic Analysis To Three Related Species’ MYB Genes Of Rose And Gene Cloning And Construction Of Over-Expression Vectors

The three plant species, Rosa chinensis, Rosa rugosa and Rosa multiflora, which are belong to the genus Rosa in Rosaceae, are famous ornamental plants in the world. Attributing to the beautiful flowers and high commerial value, the rose has been widely cultivated in China. R. rugosa and R. multiflora also have important economic value. Although the genetic relationship among the three species are close, there is significant different in morphology and metabolites in particular organs. Until now the studies about the rose have been most focused on plant regeneration and genetic transformation, while the related research about flower color and scent was little documented.In our study, the transcriptome data of the three species were analyzed by bioinformatics method. Comparative analysis were performed to their MYB transcription factors, here via compairing to the relusts by phylogenetic and expression analysis in model species. We selected the full length MYB genes from the transcriptome data, designed primers to amplified the sequences containing the complete CDS from the rose genomic DNA or cDNA library. And finally, we used these rose MYB gene clones to construct the over-expression vectors to further function identification. The main results are as follows:1. A genome-wide analysis, identified at least198MYB genes, was undertaken in strawberry and we unified renamed the MYB gene family in the species. Gene structure and chromosomal distribution comparative analysis revealed that the MYB family genes possess unequal number of introns, ranging from0to19. The distribution and density of the MYB genes on the seven chromosomes were not uniform. Strawberry chromosome6and5contained highest number of MYB genes and both ends of chromosomes are rich in MYB genes.2. We obtained79distinct full length MYB genes from the three species transcriptome data. Among them, there were34MYB genes belonged to1R type. The differential expression analysis of these79MYB genes between any two species of the genus Rosa suggested that there were at least37MYB genes expressing significantly different. These79MYB genes were employed to the phylogenetic analysis with strawberry and Arabidopsis MYB gene family.3. We screened five full length MYB genes from the rose transcriptome data, and amplified these genes using the rose genomic DNA or cDNA library as a template. Two of these five genes, RrMYB2and RrMYB10, were classified to1R type MYB, and the remaining three genes, RrMYB3, RrMYB6and RrMYB8, were belonged to the R2R3-MYB. Among them, the genomic DNA was used as template for RrMYB8, and the rose cDNA library for other cloned genes.4. We successfully constructed five over-expression vectors of rose MYB genes. They were pCAMBIA2300s-RrMYB2, pCAMBIA2300s-RrMYB3, PMV-RrMYB6, pCAMBIA2300s-RrMYB8and pCAMBIA2300s-RrMYB10vectors respectively. These five recombinant plasmids were transformed into agrobacterium EHA105.